Development of a VHH-Based Erythropoietin Quantification Assay

Molecular Biotechnology
Stefan KolPim Hermans

Abstract

Erythropoietin (EPO) quantification during cell line selection and bioreactor cultivation has traditionally been performed with ELISA or HPLC. As these techniques suffer from several drawbacks, we developed a novel EPO quantification assay. A camelid single-domain antibody fragment directed against human EPO was evaluated as a capturing antibody in a label-free biolayer interferometry-based quantification assay. Human recombinant EPO can be specifically detected in Chinese hamster ovary cell supernatants in a sensitive and pH-dependent manner. This method enables rapid and robust quantification of EPO in a high-throughput setting.

References

Sep 18, 2009·Combinatorial Chemistry & High Throughput Screening·Joy ConcepcionHong Tan
May 3, 2011·Human Molecular Genetics·Robert M Kotin
Jul 1, 2011·Current Opinion in Biotechnology·Ykelien L Boersma, Andreas Plückthun
Jul 24, 2012·Applied Microbiology and Biotechnology·Jae Seong LeeGyun Min Lee
Mar 19, 2013·Annual Review of Biochemistry·Serge Muyldermans
Feb 12, 2014·Methods in Molecular Biology·Pim HermansFrank Detmers
Oct 10, 2014·Nature Biotechnology·Gary Walsh

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Citations

Dec 15, 2015·Scientific Reports·Henning Gram HansenHelene Faustrup Kildegaard
Aug 1, 2018·ACS Synthetic Biology·Lise Marie GravHelene Faustrup Kildegaard
Feb 27, 2019·ACS Synthetic Biology·Nuša PristovšekHelene Faustrup Kildegaard

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Datasets Mentioned

BETA
DB00073

Methods Mentioned

BETA
ELISA
biosensor
PCR
Transfection
biosensors
gel filtration
affinity purification
glycosylation
surface plasmon resonance

Software Mentioned

GeneArt
Octet System Data Analysis

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